2D nightmare

Chi-Kuang Wen cwen at aux.btny.purdue.edu
Sat Jun 8 20:36:41 EST 1996


In article <199606080501.BAA29479 at gramercy.ios.com>, jgluz at STUD.MED.CORNELL.EDU wrote:
>I am trying to run the first dimension of a 2D gel using the Biorad mini 
>gel system.  I cannot get them to run.  At first, they run at about 750V 
>with a current of about 10 mAmps.  Eventually, the resistance gets so 
>high they just won't run at all (about thirty minutes).  No matter what 
>power supply I use, the current slowly drifts down to 1 mAmp and then 
>the power supply goes to fault.  I've gotten all kinds of explanations 
>from Biorad but none is satisfactory.  Has anyone ever gotten this thing 
>to work?  All advice, specific and general, is welcome.                        
>         
>--jgluz at stud.med.cornell.edu  
Hi,
Supposedly, if every condition you have used is right, you should observe a 
decrease in the current conductivityafter the proteins have reached their pIs. 
 At this point, the proteins can not conduct the current.
Therefore you encountered a very high resistance, according to V=IR, where V 
is voltage, I is current and R is resistance.  You should set constant 
amperage to run the IEF gel, but not constant voltage.   

best wishes,

Chi-Kuang Wen
>

Chi-Kuang Wen
Botany & Plant Pathology
Purdue University
West Lafayette, IN 47907
USA

fax(work): (317)494-5896
phone(work):(317)494-0661


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